An experimental forest ecosystem drought: Drought is affecting many of the world’ s forested ecosystems, but it has proved challenging to develop an ecosystem-level mechanistic understanding of the ways that drought affects carbon and water fluxes through forest ecosystems. Werner et al . used an experimental approach by imposing an artificial drought on an entire enclosed ecosystem: the Biosphere 2 Tropical Rainforest in Arizona (see the Perspective by Eisenhauer and Weigelt). The authors show that ecosystem-scale plant responses to drought depend on distinct plant functional groups, differing in their water-use strategies and their position in the forest canopy. The balance of these plant functional groups drives changes in carbon and water fluxes, as well as the release of volatile organic compounds into the atmosphere. —AMS

Temperature (T) and vapour pressure deficit (VPD) are important drivers of plant hydraulic conductivity, growth, mortality, and ecosystem productivity, independently of soil water availability. Our goal was to disentangle the effects of T and VPD on plant hydraulic responses. Young trees of Fagus sylvatica L., Quercus pubescens Willd. and Quercus ilex L. were exposed to a cross‐combination of a T and VPD manipulation under unlimited soil water availability. Stem hydraulic conductivity and leaf‐level hydraulic traits (e.g., gas exchange and osmotic adjustment) were tracked over a full growing season. Significant loss of xylem conductive area (PLA) was found in F. sylvatica and Q. pubescens due to rising VPD and T, but not in Q. ilex. Increasing T aggravated the effects of high VPD in F. sylvatica only. PLA was driven by maximum hydraulic conductivity and minimum leaf conductance, suggesting that high transpiration and water loss after stomatal closure contributed to plant hydraulic stress. This study shows for the first time that rising VPD and T lead to losses of stem conductivity even when soil water is not limiting, highlighting their rising importance in plant mortality mechanisms in the future.

Summary: The strong covariation of temperature and vapour pressure deficit (VPD) in nature limits our understanding of the direct effects of temperature on leaf gas exchange. Stable isotopes in CO₂ and H₂O vapour provide mechanistic insight into physiological and biochemical processes during leaf gas exchange. We conducted combined leaf gas exchange and online isotope discrimination measurements on four common European tree species across a leaf temperature range of 5–40°C, while maintaining a constant leaf‐to‐air VPD (0.8 kPa) without soil water limitation. Above the optimum temperature for photosynthesis (30°C) under the controlled environmental conditions, stomatal conductance (g_s) and net photosynthesis rate (A_n) decoupled across all tested species, with g_s increasing but A_n decreasing. During this decoupling, mesophyll conductance (cell wall, plasma membrane and chloroplast membrane conductance) consistently and significantly decreased among species; however, this reduction did not lead to reductions in CO₂ concentration at the chloroplast surface and stroma. We question the conventional understanding that diffusional limitations of CO₂ contribute to the reduction in photosynthesis at high temperatures. We suggest that stomata and mesophyll membranes could work strategically to facilitate transpiration cooling and CO₂ supply, thus alleviating heat stress on leaf photosynthetic function, albeit at the cost of reduced water‐use efficiency.

Summary: The impact of climate extremes on forest ecosystems is poorly understood but important for predicting carbon and water cycle feedbacks to climate. Some knowledge gaps still remain regarding how drought‐related adjustments in intra‐annual tree‐ring characteristics directly impact tree carbon and water use. In this study we quantified the impact of an extreme summer drought on the water‐use efficiency and carbon sequestration of four mature Norway spruce trees. We used detailed observations of wood formation (xylogenesis) and intra‐annual tree‐ring properties (quantitative wood anatomy and stable carbon isotopes) combined with physiological water‐stress monitoring. During 41 d of tree water deficit, we observed an enrichment in ¹³C but a reduction in cell enlargement and wall‐thickening processes, which impacted the anatomical characteristics. These adjustments diminished carbon sequestration by 67% despite an 11% increase in water‐use efficiency during drought. However, with the resumption of a positive hydric state in the stem, we observed a fast recovery of cell formation rates based on the accumulated assimilates produced during drought. Our findings enhance our understanding of carbon and water fluxes between the atmosphere and forest ecosystems, providing observational evidence on the tree intra‐annual carbon sequestration and water‐use efficiency dynamics to improve future generations of vegetation models.

Drought alters carbon (C) allocation within trees, thereby impairing tree growth. Recovery of root and leaf functioning and prioritized C supply to sink tissues after drought may compensate for drought-induced reduction of assimilation and growth. It remains unclear if C allocation to sink tissues during and following drought is controlled by altered sink metabolic activities or by the availability of new assimilates. Understanding such mechanisms is required to predict forests’ resilience to a changing climate. We investigated the impact of drought and drought release on C allocation in a 100-y-old Scots pine forest. We applied ¹³CO₂ pulse labeling to naturally dry control and long-term irrigated trees and tracked the fate of the label in above- and belowground C pools and fluxes. Allocation of new assimilates belowground was ca. 53% lower under nonirrigated conditions. A short rainfall event, which led to a temporary increase in the soil water content (SWC) in the topsoil, strongly increased the amounts of C transported belowground in the nonirrigated plots to values comparable to those in the irrigated plots. This switch in allocation patterns was congruent with a tipping point at around 15% SWC in the response of the respiratory activity of soil microbes. These results indicate that the metabolic sink activity in the rhizosphere and its modulation by soil moisture can drive C allocation within adult trees and ecosystems. Even a subtle increase in soil moisture can lead to a rapid recovery of belowground functions that in turn affects the direction of C transport in trees. Significance Climate change increases the frequency of drought events and leads to higher variability in precipitation. Drought impairs rhizosphere (root and the root-associated microbiome) functioning in trees and leads to a reduced assimilate supply belowground. It remains unclear if rhizosphere and thus whole-tree functioning can quickly recover after drought release. We show that rhizosphere metabolic activity in previously drought-exposed 100-y-old Scots pine increased in response to subtle soil moisture increases (induced by light rainfall). As a consequence of this activity change, the belowground allocation of new assimilates was immediately stimulated. Even light rainfall events can lead to a fast recovery of rhizosphere functioning and the increased C and energy demand is instantly met by altered whole-tree assimilate allocation.

Recent studies have challenged the interpretation of plant water isotopes obtained through cryogenic vacuum distillation (CVD) based on observations of a large ²H fractionation. These studies have hypothesized the existence of an H-atom exchange between water and organic tissue during CVD extraction with the magnitude of H exchange related to relative water content of the sample; however, clear evidence is lacking. Here, we systematically tested the uncertainties in the isotopic composition of CVD-extracted water by conducting a series of incubation and rehydration experiments using isotopically depleted water, water at natural isotope abundance, woody materials with exchangeable H, and organic materials without exchangeable H (cellulose triacetate and caffeine). We show that the offsets between hydrogen and oxygen isotope ratios and expected reference values (Δ²H and Δ¹⁸O) have inversely proportional relationships with the absolute amount of water being extracted, i.e. the lower the water amount, the higher the Δ²H and Δ¹⁸O. However, neither Δ²H nor Δ¹⁸O values, were related to sample relative water content. The Δ²H pattern was more pronounced for materials with exchangeable H atoms than with non-exchangeable H atoms. This is caused by the combined effect of H exchange during the incubation of materials in water and isotopic enrichments during evaporation and sublimation that depend on absolute water amount. The H exchange during CVD extraction itself was negligible. Despite these technical issues, we observed that the water amount-dependent patterns were much less pronounced for samples at natural isotope abundance and particularly low when sufficiently high amounts of water were extracted (>600 µL). Our study provides new insights into the mechanisms causing isotope fractionation during CVD extraction of water. The methodological uncertainties can be controlled if large samples of natural isotope abundance are used in ecohydrological studies.